Kamst E, Pilling J, Raamsdonk L M, Lugtenberg B J, Spaink H P
Institute of Molecular Plant Sciences, Leiden University, The Netherlands.
J Bacteriol. 1997 Apr;179(7):2103-8. doi: 10.1128/jb.179.7.2103-2108.1997.
Synthesis of chitin oligosaccharides by NodC is the first committed step in the biosynthesis of rhizobial lipochitin oligosaccharides (LCOs). The distribution of oligosaccharide chain lengths in LCOs differs between various Rhizobium species. We expressed the cloned nodC genes of Rhizobium meliloti, R. leguminosarum bv. viciae, and R. loti in Escherichia coli. The in vivo activities of the various NodC proteins differed with respect to the length of the major chitin oligosaccharide produced. The clearest difference was observed between strains with R. meliloti and R. loti NodC, producing chitintetraose and chitinpentaose, respectively. In vitro experiments, using UDP-[14C]GlcNAc as a precursor, show that this difference reflects intrinsic properties of these NodC proteins and that it is not influenced by the UDP-GlcNAc concentration. Analysis of oligosaccharide chain lengths in LCOs produced by a R. leguminosarum bv. viciae nodC mutant, expressing the three cloned nodC genes mentioned above, shows that the difference in oligosaccharide chain length in LCOs of R. meliloti and R. leguminosarum bv. viciae is due only to nodC. The exclusive production of LCOs which contain a chitinpentaose backbone by R. loti strains is not due to NodC but to end product selection by Nod proteins involved in further modification of the chitin oligosaccharide. These results indicate that nodC contributes to the host specificity of R. meliloti, a conclusion consistent with the results of several studies which have shown that the lengths of the oligosaccharide backbones of LCOs can strongly influence their activities on host plants.
NodC合成几丁质寡糖是根瘤菌脂几丁质寡糖(LCOs)生物合成中的首个关键步骤。不同根瘤菌属物种的LCOs中寡糖链长度的分布有所不同。我们将苜蓿根瘤菌、豌豆根瘤菌蚕豆生物型和百脉根根瘤菌的克隆nodC基因在大肠杆菌中进行了表达。不同NodC蛋白的体内活性在产生的主要几丁质寡糖长度方面存在差异。在分别产生几丁质四糖和几丁质五糖的苜蓿根瘤菌和百脉根根瘤菌NodC菌株之间观察到了最明显的差异。使用UDP-[14C]GlcNAc作为前体的体外实验表明,这种差异反映了这些NodC蛋白的内在特性,且不受UDP-GlcNAc浓度的影响。对表达上述三个克隆nodC基因的豌豆根瘤菌蚕豆生物型nodC突变体产生的LCOs中的寡糖链长度进行分析,结果表明苜蓿根瘤菌和豌豆根瘤菌蚕豆生物型LCOs中寡糖链长度的差异仅归因于nodC。百脉根根瘤菌菌株仅产生含有几丁质五糖主链的LCOs并非由于NodC,而是由于参与几丁质寡糖进一步修饰的Nod蛋白对终产物的选择。这些结果表明nodC对苜蓿根瘤菌的宿主特异性有贡献,这一结论与多项研究结果一致,这些研究表明LCOs寡糖主链的长度可强烈影响其对宿主植物的活性。